Composition for the treatment and/or prevention of coccidiosis in poultry



United States Patent Int. (:1. Milk 27/00 US. Cl. 424-251 18 ClaimsABSTRACT OF THE DISCLOSURE Anti-coccidiosis activity of compounds of theformula wherein Q is NH or NH +X*, each of R and R is a lower alkylgroup with from 1 to 3 carbon atoms, and each of X and X'- is aninorganic acid anion, is remarkably increased by the coemploymenttherewith of free terephthalic acid or terephthalic acid monoor diesteror of salt of the said acid or mono-ester.

The present invention relates to compositions for the treatment and/orprevention of coccidiosis in poultry, and to methods for the treatmentand/or prevention of coccidiosis in poultry.

Coccidiosis is an infectious fatal poultry disease and the treatment andprevention of the disease constitute one of the most important problemsin the poultry industry.

Attempts have heretofore been made to treat the disease by means ofvarious anti-coccidial agents such as arsenic compounds, nitrofurazone,bisphenol, sulfa drugs, pyrimidinyl methyl imidazolium salts, andpyrimidinylmethyl pyridinium salts. These known anti-coccidial drugs,however, have such defects as Weak activity or otherwise, strong andunfavorable side-effects.

We have previously found that thiazolium quaternary salts of the formulathe formula Q N s R onzi t I N WE wherein each of R and R is lower alkylwith from 1 to 3 carbon atoms, and Q, X'" and X have the previouslyrecited significances, is remarkably increased by the coemploymenttherewith of free terephthalic acid or certain terephthalic acidderivatives, as hereinafter defined.

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That is to say, the present invention provides compositions for thetreatment and/or prevention of coccidiosis in poultry, whichsubstantially consist of admixtures of carrier and/or diluent andthiazolium salt of the abovementioned Formula I and terephthalic acid orterephthalic acid derivative, the concentration of the thiazolium saltin the composition being from 0.001% to about 0.05% by weight and theconcentration of the terephthalic acid or derivatives thereof being fromabout 0.1% to about 0.5% by weight. It also provides a method oftreating and/ or preventing coccidiosis in poultry, in which ananti-coccidial amount of the thiazolium salt of the above mentionedFormula I and terephthalic acid or derivative thereof are administeredto poultry.

In Formula I, each of R and R is methyl, ethyl, propyl or isopropyl, X'"and X- stands for the same or different (advantageously the same)inorganic acid anions, e.g. Cl", Br, I NO3 NO2 C1043 SCN", etc.

Typical thiazolium quaternary salts (I) constituting the activeanti-coccidial agents in the compositions of the invention comprise, forexample:

3-(2-methyl-4-amino-pyrimidin-S-yl-methyl)-4,5-dimethylthiazolium salt;

3 2-methyl-4-arnino-pyrimidin-5-yl-methyl) -4-methy1- S-ethylthiazoliumsalt;

3 2-methyl-4-amino-pyrimidin-5-yl-methyl) -4-methyl- S-propylthiazoliumsalt, and

3 (2-ethyl-4-a-minopyrimidin-5 -yl-methyl -4, S-dimethylthiazolium salt,

(salt referring to the chloride, bromide, iodide, nitrate, nitrite,sulfate, perchlorate, thiocyanate, etc. monoor di-salt).

A compound of Formula I can be prepared e.g. by reacting a compound offormula N R TIN-H2 OH NH V 2 2 (I!) with carbon disulfide and3-halo-alkan-2-one, e.g. 3- chloro-butan-Z-one, in the presence of abasic compound such as aqueous ammonia, alkali metal hydroxide or alkalimetal carbonate to give '3-(2-loweralkyl-4-aminopyrimidin-S-yl-methyl)-4-hydroxy-4-methyl-5-lower alkylthiazolidine-Z-thione of formula N s re -f N112 H N OHz-N s on, R (III)ester, diethyl ester, dipropyl ester and dibutyl ester),

pharmaceutically acceptable terephthalic acid monoor di-salts (e.g.mono-sodium salt, mono-potassium salt, disodium salt and dipotassiumsalt), and pharmaceutically acceptable mono-salts of the mono-esters.The salts with X and X, as previously enumerated, are pharmaceuticallyacceptable for administration to poultry.

3 4 The use of free terephthalic acid is most advantageous. Acomposition according to the present invention com- Test compound, TPA,prises a thiazolium quaternary salt of Formula I and percent percentterephthalic acid or derivative thereof, as above-defined, Testcompoundgroup, as active anti-coccidial ingredients dispersed in, or ad- A0.0063 mixed with, a suitable carrier or diluent. The composi- 5 M063tions can be administered in such forms as powders, gran- (5) Method oftest: Above-mentioned chicks are raised ules, tablets, suspension,emulsion or injection or advanisolatedly for at least 7 days and thecondition of their tageously are administered in admixture with poultrydiet health observed. They are weighed and divided into four or insolution in drinking water for the poultry depending 10 groupsconsisting of five chicks each. The mean weight on various conditionssuch as the kind of poultry, the of each group is arranged to be equal.Two of the groups state of the poultry or the age of the poultry. Ascarri r are test groups and the remainder are control groups. The ordiluent, it is preferred to use one which is not reactive chicks in thetest groups are fed the diet containing the with the t fi ingredients,thialolibm Salts and composition according to this invention and thecontrol ephthalie acid or derivative thereof, and which can be groupsare fed the normal poultry diet. After 24 hours, administered withsafety to poultry. Carriers or diluents each chick except non-infectiousgroup is inoculated suitable in the present compositions include, forexample, orally with 180,000 oocyst of Eimeria. tenella. Wheat $110115,Wheat flour, corn meal, SOB/b62111 mill fe d, (6) Observations:Mortality and bloody droppings in y n Vegetables, ground shells, fishmeal, soyeach group are recorded daily until the end of the test. beangrits, dried grains, fermentation residues, rice-bran, Eight days afteroocyst inoculation, all the survivors are potato starch, lactose,sucrose, glucose, fructose, molasses sacrificed and examinedpathologically. solubles, antibiotic-containing mycelia, vitamins,crushed (7) Explanation of findings set forth in the table: limestone,clay, etc.

The optimum concentration of the present composition Increasmg rate ofbody Welght depends to some extent on the kind of thiazolium salt 25Average body weight at the Finish 1 100 employed, the kind of poultry orthe purpose of the ad- Average body weight at the Start ministration ofthe composition, that is prevention or Rate of increasing rate of bodyweight: curing. A composition of any desired concentration may beprepared by selecting proper diluents and by determinlncreefsmg rate ofbody i of test group X 00 ing the proper ratio of carrier to activeingredients. In Increasmg rate f body welght of control group general,compositions containing from about 0.001% to Bloody droppings: Averagedrops of blood excreted about 0.05% by weight, preferably from about 0.05% from the intestine of each chick belonging to a group are to about0925% y Wright of tbialolillm Salt and shown according to the results ofobservation. containing about from 01% about 05%, Preferably Severity ofcaecal lesions (shown b the following symfrom about 01% t0 03%, y Weightof tefepbtbalic acid bols according to the results of observation): orderivative thereof in various poultry diets or water are N m 1 lsuitable lflor the treatment and the prevention of the disf no eslonsease. W en the thiazolium salts (I) are employed as therapeutic agents,a higher concentration (about 0.0125 fii P 10 3 g t to 0.05%) ispreferably used and when the thiazolium 40 0 f eslons mar ut remeSevere, caeca greatly distended with blood or salts (I) are employed aspreventive agents, a lower con- 1 n 1 k centration 0.003%0.0l25% ispreferably used. caseous core caaca. Wa um orm y very c Intermediateformulations of a certain concentration Oocyst output (0.P.G.): Numberof oocysts in 1 gram are preferably prepared in advance by usingsuitable diof caecal contents. luents or carriers, and added to thefinished feed in ad- Rate of sporulwtion: To the caecal contents, 2%aqueministration to poultry in the required proportions. About oussolution of potassium bichromate, the volume being 1% to 30% by weightof the active ingredients is con- 10 times as much as the contents, isadded. Then the convenient as the concentration of the intermediateformulatents are incubated at 28 C. for 3, days, and the pertions.centage of the sporulated oocysts to the all in the content Thefollowing illustrative experiments are given for a is calculated. betterunderstanding of the invention. Result: The result is shown in thetable.

TABLE Rate of Oocysts l ncreas- 11101038- Average ing rate ing rateBlood droppings atter- Severity of cocoa lesions Rate of body of body orbody M (percent Oocyst sporu- Percent by weight (of com- Weight weight,weight, Mor- 4 5 6 7 output lation, poundindiet) at start percentpercent tality days days days days t X104 percent Test group:

A(Test cpd., 0.00s3%) 54.4 150.0 110.0 o/s 0.4 0.6 7/10 3/10 80 12.5 B(Test cpcL, 0.0003% plus TPA,0.5%) 54.4 151.0 110.8 0/5 2/10 8/10 10 0Control group:

I(Infectious control) 54.4 104.8 76.8 7/15 1.8 m 1.7 14/16 2/16 500 82.II (Noniufectious control) 54.4 136.4 100.0 O/5 10/10 0 0 TEST RUN 1Test procedures (1) Chicks: Nine-day-old white leghorn males.

-(2) Oocyst: Sporulated oocysts of Eimeria tenella.

(3) Administered compounds: 3 (2 methyl 4- aminopyrimidin 5 yl methyl)4,5 dimethylthiazolium chloride hydrochloride (test compound) andterephthalic acid (TPA).

(4) Mixture rate: The run is carried out by employing starter feeds Aand B respectively containing test compound and/ or TPA as follows:

As shown in the table, the anti-coccidial action of the 3 (2 methyl 4aminopyrimidin 5 yl methyl) 4,5- dimethylthiazolium salt is increased bythe coemployment of terephthalic acid therewith. Similiar enhancement isachieved also with the other thiazolium salts.

In the following illustrative examples, part or parts are shown on aweight basis, unless otherwise specified. Parts by Weight bear the samerelation to parts by volume as do grams to milliliters.

EXAMPLE 1 Preparation of active ingredient 7.5 parts of2-methyl-4-amino-5-aminomethylpyridine is dissolved in 40 parts byvolume of 80% methanol. To the solution, there are added 3.4 parts of28% aqueous ammonia, 4.2 parts of carbon disulfide and then 8.1 parts of3-chlorobutan-2-one, and the mixture is agitated for 4 hours at 50 C.,reaction taking place. After the reaction, the methanol is distilled offunder reduced pressure and to the residue is added 7.2 parts ofconcentrated hydrochloric acid, followed by heating at 70 C. for 1 hour.After cooling, an aqueous solution of sodium hydroxide is added to thesolution and the pH value is adjusted to 7, solid substance being throwndown. The solid substance is separated by filtration, washed with waterand dried to give 13 parts of 3-(2-methyl-4-aminopyrimidin-5-yl-methyl)-4,5-dimethylthiazole-2-thione melting at 223 C. Yield 90%.

parts of 3-(2-methyl-4-arninopyrimidin-S-yl-methyl)-4,S-dimethylthiazole-Z-thione is dissolved in 100 parts by volume ofWater. To the solution there is Slowly added 10.2 parts of 30% hydrogenperoxide under agitation while the temperature is kept at 20 C. Theresulting mixture is agitated for 2 hours to allow a reaction to takeplace. 5.5 parts of sodium bicarbonate is added to reaction mixture toadjust the pH value to the neutral range, followed by adding to theso-treated total mixture a concentrated aqueous solution of 2.5 parts ofammonium thiocyanate to give 8.0 parts by weight of 3-(2-methyl-4-aminopyrimidin 5 yl methyl) 4,5 dimethylthiazolium thiocyanate. (Yield80%) Ml. 188-190 C.

Elementary analysis-Calculated for C H N S .H O: C, 46.28; H, 5.50; N,22.49. Found: C, 45.89; H, 5.36; N, 22.21.

In place of the above ammonium thiocyanate a concentrated aqueoussolution of 2.5 parts by weight of sodium nitrate is employed to give6.5 parts of 3-(2-methyl-4-aminopyrimidin-S-yl-methyl)-4,5-dimethylthiazolium nitrate. (Yield 68%.)

After recrystallizing from hot water, the nitrate-is in the form ofcolorless needles melting at 185-188 C. (decomposition).

Elementary analysis.-Calculated for C I-I N O S: C, 44.43; H, 5.08; N,23.55. Found: C, 44.26; H, 5.20; N, 23.35.

10 parts of 3-(2-methyl-4-aminopyrimidin-S-yI-methyl)-4,5-dimethylthiazole-2-thione is suspended in 40 parts by volume ofwater, followed by adding 9.2 parts by Weight of dihydrate of bariumchloride. To the suspension, there is slowly added 11.6 parts by volumeof 30% hydrogen peroxide under agitation while the temperature is keptat C. The resulting mixture is agitated for 3 hours to allow a reactionto take place. After the reaction, barium sulfate is filtered off. Thefiltrate is concentrated under reduced pressure and 100 parts by volumeof ethanol added thereto, followed by cooling to give crystals. Thecrystals are collected and dried to give 9.4 parts of 3-(2-methyl-4-aminopyrimidin-S-yl-methyl)-4,5-dimethyl thiazolium chloridehydrochloride as colorless needles melting at 232 C. (decomposition).Yield 87%.

The other salts of this invention are prepared in analogous manner.

EXAMPLE 2 Formulations (A) The following components are admixed:

Parts 3 (2 methyl 4 aminopyrimidin 5 yl methyl)- 4,5-dimethylthiazoliumchloride hydrochloride 0.3 Terephthalic acid 24.7 Wheat flour 75 1 partof the mixture is homogeneously added to 99 parts of conventional dietfor chicks.

Thus-prepared composition is fed to chicks. (B) The following componentsare admixed:

Parts 3-(2 me'thyl 4 aminopyrimidin 5 yl methyl),

4,5-dimethyl-thiazolium chloride hydrochloride 0.1

Terephthalic acid 9.9 Soybean flour 85 Sucrose 5 2 parts of the mixtureis homogeneously added to 98 parts of conventional poultry diet forbreeders.

Thuswprepared composition is fed to breeders. (D) The followingcomponents are admixed:

Parts 3 (2 methyl 4 aminopyrimidin 5 yl methyl)- 4,5-dimethyl-thiazoliumchloride hydrochloride 0.1

Terephthalic acid 5.9 Lactose 79 Sucrose 15 5 parts of the mixture isdissolved in 100 parts by volume of water.

Thus-prepared composition is fed to chicks. (E) The following componentsare admixed:

Parts 3-(2 methyl 4 aminopyrimidin 5 yl methyl)- 4,5-dimethyl-thiazoliumchloride hydrochloride 0.01

Terephthalic acid 0.99 Grain flour 88 Calcium carbonate 11 50 parts ofthe mixture is homogeneously added to 50 parts of conventional poultrydiet for layers.

Thus-prepared composition is fed to layers. (F) The following componentsare admixed:

Parts 3-(2 methyl 4 aminopyrimidin 5 yl methyl)- 4,5-dimethyl-thiazoliumthiocyanate 0.3 Terephthalic acid 24. Wheat flour One part of themixture is homogeneously added to 99 parts of conventional diet forchicks.

Thus-prepared composition is fed to chicks. (G) The following componentsare admixed:

Parts 3 (2 methyl 4 aminopyrimidin 5 yl methyl)-4,5-dimethyl-thiazoliurn nitrate 0.1 Terephthalic acid 9.9 Soybean flour85 Sucrose 5 Two parts of the mixture is homogeneously added to 98 partsof conventional poultry diet for breeders.

Thus-prepared composition is fed to breeders. (H) The followingcomponents are admixed:

Parts 3-(2 methyl 4 aminopyrimidin 5 yl methyl)- 4,5-dimethyl-thiazoliumthiocyanate 0.2 Terephthalic acid 19.8 Fermentation residue Two parts ofthe mixture is homogeneously added to 98 parts of conventional poultrydiet for breeders.

Thus-prepared composition is fed to breeders.

Five parts of the mixture is dissolved in 100 parts by volume of water.

Thus-prepared composition is fed to chicks.

(J) The following components are admixed:

. Parts 3 (2 methyl 4 aminopyrimidin yl-methyl)- 4,5-dimethyl thiazoliumnitrate 0.01 Terephthalic acid 0.99 Wheat flour 88 Calcium carbonate 1125 parts of the mixture is homogeneously added to 75 parts ofconventional poultry diet for layers.

Thus-prepared composition is fed to layers. (K) The following componentsare admixed:

Parts 3 (2 methyl 4 aminopyrimidin 5 yl-methyl)- 4,5-dimethyl-thiazoliummononitrate 0.3 Terephthalic acid 24. Wheat flour 75 1 part of themixture is homogeneously added to 99 parts of conventional diet forchicks.

Thus-prepared composition is fed to chicks.

Feeding poultry with any of the above-described compositions iseffective in preventing and/or in alleviating coccidiosis.

Although the present invention has been described in conjunction withpresently preferred embodiments, essentially the same procedures can befollowed with any of the hereindescribed compositions, with essentiallysimilar results.

If the effective amount, i.e. the anti-coccidial amount, of thethiazolium salt to be administered according to the present invention iscalculated in terms of the amount per 100 grams of the body weight ofpoultry, it may also be expressed as about 0.3 to 20 milligrams,preferably about 1 to milligrams, per 100 grams of the body weight ofpoultry, and amount of terephthalic acid may be preferably expressed asabout 30 to about 170 milligrams, preferably about 60 to 100 milligrams,per 100 grams of the body weight of poultry.

What is claimed is:

1. A method of treating or preventing coccidiosis in poultry whichcomprises administering to the poultry an anti-coccidial amount of anadmixture of (a) thiazolium salt of the formula Q Rt N hair wherein eachof R and R is alkyl with a maximum of 3 carbon atoms, Q is a member of-NH and NH +X each of X and X- is an inorganic acid ion, and (b) amember selected from the group consisting of free'terephthalic acid,terephthalic acid monoand di-alkyl esters with a maximum of 4 carbonatoms in an alkyl group and pharmaceutically acceptable salts of saidacid and monoesters, and (c) an ingestible pharmaceutical carrier (a)and (b), the concentration of component (a) in the total compositionbeing from about 0.001% to about 0.05% by weight and the concentrationof component (b) in the total composition being from about 0.1% to about0.5% by weight. I

2. A method according to claim 1 wherein the pharmaceutically acceptablesalts are sodium or potassium salts.

3. A method fit t tu'ding to claim 1 wherein X and 8 X- areindependently selected from the group consisting of anions Cl-, Br-, I-,N0 N0 1 (SO. /2, C10; and SCN. 1

4. A method according to claim 1 wherein X- and X'* are independentlyselected from the group consisting of anions Cl-, Br, 1*, N0 1 NO (SO/2, C10 and SCN, and'the pharmaceutically acceptable salts are sodium orpotassium salts.

5. The method according to claim 1, wherein (a) is 3 (2 methyl 4aminopyrimidin 5 yl methyl)- 4,5-dimethylthiazolium choridehydrochloride and (b) terephthalic acid.

' 6. The method according to claim 1, wherein (a) is 3 (2 methyl 4 aminopyrimidin 5 yl methyl)- -4,5-dimethylthiazolium nitrate and (b)terephthalic acid.

7. The method according to claim 1, wherein (a) is 3 (2 methyl 4 aminopyrimidin 5 yl methyl)- 4,5-dimethylthiazolium thiocyanate and (b) isterephthalic acid. 1

8. The method according to claim 1, wherein (a) is 3 *(2 ethyl 4 aminopyrimidin 5 yl methyl)- 4,5-dimethylthiazoliurn nitrate and (b) isterephthalic acid.

9. A composition for the treatment or prevention of coccidiosis inpoultry which comprises an ingestible pharmaceutical carrier and as itsactive ingredients, (a) thiazolium salt of the formula wherein each of Rand R is alkyl with a maximum of 3 carbon atoms, Q is a member of NH and--NH X'-, each of X" and X' is an inorganic acid anion, and (b) a memberselected from the group of free terephthalic acid, terephthalic acidmonoand di-alkyl esters with a maximum of 4 carbon atoms in an alkylgroup and pharmaceutically acceptable salts of said acid andmono-esters, the concentration of said active ingredient (a) in thetotal composition being from 0.001% to about 0.05% by weight and theconcentration of said active ingredient (b) in the total compositionbeing from about 0.01% to about 0.5% by weight.

10. A composition according to claim 9 wherein the pharmaceuticallyacceptable salts are sodium or potassium salts.

11. A composition according to claim 9 wherein X" and X' areindependently selected from the group consisting of anions Cl, Br, I-,N0 1 N0 (SO /2, C10; and SCN.

12. A composition according to claim 9 wherein X and X' areindependently selected from the group consisting of anions Cl-, BY", 1*,N0 1 N0 (SO /2, C10 and SCN, and the pharmaceutically acceptable saltsare sodium or potassium salts.

13. A composition according to claim 9, wherein (a) is 3 (2 methyl 4aminopyrimidin 5 yl methyl)- 4,5-dimethylthiazolium chloridehydrochloride and (b) is terephthalic acid.

14. A composition according to claim 9, wherein (a) is 3 (2 methyl 4aminopyrimidin 5 yl methyl)- 4,5 -dimethy1thiazolium nitrate and (b) isterephthalic acid.

15. A composition according to claim 9, wherein (a) is 3 (2 methyl 4aminopyrimidin 5 yl methyl)- 4,5-dimethylthiazolium thiocyanate and (b)is terephthalic acid.

16. A composition according to claim 9, wherein (a) is 3 (2. ethyl 4aminopyrimidin 5 yl methyl)- 4,5-dimethylthiazsolium nitrate and (b) isterephthalic acid.

17. A composition according to claim 9, wherein the carrier or diluentis essentially constituted by poultry feed.

9 10 18. A composition according to claim 9, wherein the OTHERREFERENCES carrier or diluent is essentially constituted by drinkingVeterinary Bulletin, 4138, 1961. Water the Pwltry- Veterinary Builietin,No. 83, 1960.

References Cited 5 HOWARD M. ELLIS, Primary Examiner UNITED STATESPATENTS FRANK CACCIAPAGLIA, 111., Assistant Examiner 2,855,340 9/1958Kent et a1. 16753 3,268,403 8/1966 Rogers et a1. 16753.1

3,304,227 2/1967 Loveless 167--53.1 10 424-408, 317

